title: Evidence for the suppression of apoptosis by the peroxisome proliferator activated receptor alpha (PPAR alpha).
abstract: Peroxisome proliferators (PPs) are a class of nongenotoxic rodent hepatocarcinogens. We have demonstrated previously that PPs suppress both spontaneous rat hepatocyte apoptosis and that induced by exogenous stimuli such as transforming growth factor-beta1 (TGFbeta1). PPs transcriptionally activate the peroxisome proliferator activated receptor-alpha (PPAR alpha), a member of the nuclear hormone receptor superfamily. Here, we investigate whether activation of PPAR alpha mediates the suppression of rat hepatocyte apoptosis induced by PPs. We isolated a naturally occurring variant form of PPAR alpha (hPPAR alpha-6/29) from human liver by PCR cloning. Electrophoretic mobility shift assays (EMSA) demonstrated that hPPAR alpha-6/29 shared the ability of mPPAR alpha to heterodimerise with the retinoid X receptor (RXR) and bind to DNA. When hPPAR alpha-6/29 was transfected into Hepa1c1c7 cells together with a reporter plasmid containing a PPAR response element (PPRE), hPPAR alpha-6/29, unlike mPPAR alpha, could not be activated by PPs. Furthermore, hPPAR alpha-6/29 could act as a dominant negative regulator of PPAR-mediated gene transcription since increasing concentrations of hPPAR alpha-6/29 abrogated the activation of co-transfected mPPAR alpha. When introduced into primary rat liver cell cultures by transient transfection, hPPAR alpha-6/29 prevented the suppression of hepatocyte apoptosis by the PP nafenopin, but not that seen in response to phenobarbitone (PB), a nongenotoxic carcinogen whose action does not involve PPAR alpha. The suppression of hepatocyte apoptosis was abrogated completely even though only 30% of hepatocytes were transfected, suggesting the involvement of a soluble factor. These data indicate that activation of rat liver PPAR alpha provides a survival signal for hepatocytes, preventing their death in response to apoptotic stimuli.
authors.affiliation: Zeneca Central Toxicology Laboratory, Alderley Park, Macclesfield, UK. ruth.roberts@CTL.ZENECA.com
authors.forename: R A
authors.initials: RA
authors.lastname: Roberts
authors.forename: N H
authors.initials: NH
authors.lastname: James
authors.forename: N J
authors.initials: NJ
authors.lastname: Woodyatt
authors.forename: N
authors.initials: N
authors.lastname: Macdonald
authors.forename: J D
authors.initials: JD
authors.lastname: Tugwood
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular; D002478:Cells, Cultured; D017931:DNA Primers; D017930:Genes, Reporter; D006801:Humans; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D008113:Liver Neoplasms; D008233:Lymphotoxin-alpha / Q000494:pharmacology*; D008297:Male; D051379:Mice; D009687:Nuclear Proteins / Q000502:physiology; D016133:Polymerase Chain Reaction; D051381:Rats; D017208:Rats, Wistar; D018160:Receptors, Cytoplasmic and Nuclear / Q000096:biosynthesis / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D014157:Transcription Factors / Q000096:biosynthesis / Q000502:physiology*; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D017931:DNA Primers; D008233:Lymphotoxin-alpha; D009687:Nuclear Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D011994:Recombinant Proteins; D014157:Transcription Factors